Deubiquitinase USP19 in TDP-43 pathogenesis.

TDP-43 发病机制中的去泛素酶 USP19。

• 批准号: 10463231
• 负责人: David E Kang
• 金额: $ 178.52万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10463231
• 关键词: ADP ribosylation; Acetylation; Age; Aging; Alternative Splicing; Amyotrophic Lateral Sclerosis; Autophagocytosis; Autopsy; Behavioral; Biochemical; Biological; Biological Assay; Brain; Cells; Cognitive; Cohort Studies; Coupled; Cytoplasm; Cytosol; Deubiquitination; Ectopic Expression; Electrophysiology (science); Endoplasmic Reticulum; Enzymes; Family; Frontotemporal Lobar Degenerations; Functional disorder; Gene Dosage; Genetic; Hippocampus (Brain); Human; Human Genome; Image; Impaired cognition; Impairment; In Vitro; Late Onset Alzheimer Disease; Link; Lysine; Lysosomes; Mediating; Modification; Molecular; Motor; Motor Neurons; Mus; Neurons; Nuclear Protein; Outcome; Parkin; Pathogenesis; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Phenotype; Phosphorylation; Physiology; Post-Translational Protein Processing; Protein Isoforms; Proteins; Proteomics; RNA Binding; Recombinant Proteins; Regulation; Role; Severities; Site; Slice; Solubility; Sumoylation Pathway; Synapses; Synaptic plasticity; System; Testing; Therapeutic; Tissues; Toxic effect; Transcript; USP8 gene; Ubiquitin; Ubiquitination; Variant; Viral; adenoviral-mediated; age related; endoplasmic reticulum stress; human model; in vivo; insight; member; motor impairment; mouse model; multicatalytic endopeptidase complex; mutant; neuroinflammation; neurotoxicity; non-demented; novel; oxidation; protein TDP-43; protein complex; sciatic nerve; ubiquitin-protein ligase

The RNA-binding nuclear protein TDP-43 mislocalizes to the cytoplasm and aggregates in Frontotemporal lobar degeneration (FTLD-TDP variant), Amyotrophic Lateral Sclerosis (ALS), and >50% of late-onset Alzheimer’s disease (AD). Abnormal TDP-43 mislocalization and accumulation is associated with endoplasmic reticulum (ER) stress, synaptic dysfunction, and cognitive and motor impairments. While TDP-43 undergoes different post-translational modifications including phosphorylation, poly ADP-ribosylation, oxidation, acetylation, sumoylation, and ubiquitination, ubiquitination is a final key modification required for the turnover of TDP-43 via the ubiquitin-proteasome system and autophagy-lysosome pathways. TDP-43 is ubiquitinated by E3 ligases Parkin, PJA1, and Znf179. However, the role of deubiquitinases (DUBs) in the regulation of TDP-43 function, turnover, proteinopathy, and toxicity is poorly understood. The human genome encodes ~90 DUBs. Ubiquitin specific peptidases (USPs) are the largest family of DUBs comprising ~50 members in humans. Of these, 27 are expressed in the CNS. Our results from an unbiased screen of CNS-expressed DUBs identified USP19 as a major TDP-43 DUB, a positive regulator of TDP-43 stability, and a promising candidate for further study. Specifically, preliminary studies indicate that USP19, a DUB elevated during aging and in brains of FTLD-TDP patients, acts to increase TDP-43 stability/aggregation and participates in TDP-43-induced ER stress. By taking advantage of mouse models and human postmortem tissues together with molecular, cell biological, imaging, biochemical, proteomics, electrophysiological, behavioral, viral, histochemical, and recombinant protein toolsets, this proposal will 1. validate the role of USP19 in TDP-43 pathogenesis and associated phenotypes vivo, and 2. determine the mechanistic basis of USP19 in TDP-43 deubiquitination, stability, aggregation, and toxicity in genetically modified neurons and in vitro systems. Successful conclusion of these studies will determine the significant contribution of USP19 and its DUB activity to TDP-43 pathogenesis in humans and mice. Moreover, these results will provide novel mechanistic insights to USP19 DUB activity in concert with TDP-43 in ER stress and neurotoxicity. Together, these studies will enable the pursuit of a potential therapeutic direction of targeting USP19-mediated mechanisms to mitigate TDP-43 pathology and toxicity.

RNA 结合核蛋白 TDP-43 错误定位到细胞质并聚集在 额颞叶变性（FTLD-TDP 变异）、肌萎缩侧索硬化症 (ALS)、 > 50% 的迟发性阿尔茨海默病 (AD) 异常 TDP-43 定位错误和 积累与内质网 (ER) 应激、突触功能障碍和 而 TDP-43 则经历不同的翻译后障碍。 修饰包括磷酸化、聚 ADP-核糖基化、氧化、乙酰化、 sumoylation 和泛素化，泛素化是营业额所需的最后一个关键修饰 TDP-43 通过泛素-蛋白酶体系统和自噬-溶酶体途径。 被 E3 连接酶 Parkin、PJA1 和 Znf179 泛素化，但去泛素化酶的作用。 (DUB) 对 TDP-43 功能、周转、蛋白质病和毒性的调节作用很差 人类基因组编码约 90 个泛素特异性肽酶 (USP)。 最大的 DUB 家族，由约 50 个成员组成，其中 27 个在人类中表达。 我们对 CNS 表达的 DUB 进行公正的筛选，结果将 USP19 确定为 主要 TDP-43 DUB，TDP-43 稳定性的正调节剂，并且是进一步研究的有希望的候选者 具体来说，初步研究表明 USP19（一种 DUB）在衰老过程中和在体内升高。 FTLD-TDP 患者的大脑，可增加 TDP-43 的稳定性/聚集并参与 TDP-43 诱导的 ER 应激。 通过利用小鼠模型和人类死后组织以及分子， 细胞生物学、成像、生物化学、蛋白质组学、电生理学、行为学、病毒学、 组织化学和重组蛋白工具集，该提案将 1. 验证 USP19 的作用 2. 确定 TDP-43 发病机制和体内相关表型，并确定其机制基础 USP19 在 TDP-43 去泛素化、稳定性、聚集和转基因毒性中的作用 神经元和体外系统。 这些研究的成功结束将确定 USP19 和 此外，这些结果将提供其 DUB 活性对人类和小鼠 TDP-43 发病机制的影响。 对 USP19 DUB 活性与 TDP-43 在 ER 应激和 总之，这些研究将有助于寻找潜在的治疗方向。 靶向 USP19 介导的机制以减轻 TDP-43 病理学和毒性。

项目成果

The multifaceted functions of β-arrestins and their therapeutic potential in neurodegenerative diseases.

β-抑制素的多方面功能及其在神经退行性疾病中的治疗潜力。

• 发表时间: 2024-02
• 影响因子: 12.8
• 作者: Kee, Teresa R;Khan, Sophia A;Neidhart, Maya B;Masters, Brianna M;Zhao, Victoria K;Kim, Yenna K;McGill Percy, Kyle C;Woo, Jung
• 通讯作者: Woo, Jung